title: Preparation and diagnostic utility of a hemagglutination inhibition test antigen derived from the baculovirus-expressed hemagglutinin-neuraminidase protein gene of Newcastle disease virus.
abstract: A recombinant hemagglutinin-neuraminidase (rHN) protein from Newcastle disease virus (NDV) with hemagglutination (HA) activity was expressed in Spodoptera frugiperda cells using a baculovirus expression system. The rHN protein extracted from infected cells was used as an antigen in a hemagglutination inhibition (HI) test for the detection and titration of NDV-specific antibodies present in chicken sera. The rHN antigen produced high HA titers of 2(13) per 25 μL, which were similar to those of the NDV antigen produced using chicken eggs, and it remained stable without significant loss of the HA activity for at least 12 weeks at 4°C. The rHN-based HI assay specifically detected NDV antibodies, but not the sera of other avian pathogens, with a specificity and sensitivity of 100% and 98.0%, respectively, in known positive and negative chicken sera (n = 430). Compared with an NDV-based HI assay, the rHN-based HI assay had a relative sensitivity and specificity of 96.1% and 95.5%, respectively, when applied to field chicken sera. The HI titers of the rHN-based HI assay were highly correlated with those in an NDV-based HI assay (r = 0.927). Overall, these results indicate that rHN protein provides a useful alternative to NDV antigen in HI assays.
authors.affiliation: Avian Diseases Division, Animal and Plant Quarantine Agency, Anyang 430-757, Korea.kchoi0608@korea.kr.
authors.forename: Kang-Seuk
authors.initials: KS
authors.lastname: Choi
authors.forename: Soo-Jeong
authors.initials: SJ
authors.lastname: Kye
authors.forename: Woo-Jin
authors.initials: WJ
authors.lastname: Jeon
authors.forename: Mi-Ja
authors.initials: MJ
authors.lastname: Park
authors.forename: Saeromi
authors.initials: S
authors.lastname: Kim
authors.forename: Hee-Jung
authors.initials: HJ
authors.lastname: Seul
authors.forename: Jun-Hun
authors.initials: JH
authors.lastname: Kwon
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism; D016367:Baculoviridae / Q000235:genetics; D002645:Chickens; D015707:HN Protein / Q000235:genetics / Q000378:metabolism; D006385:Hemagglutination Inhibition Tests / Q000379:methods* / Q000662:veterinary; D009521:Newcastle Disease / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D009522:Newcastle disease virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011201:Poultry Diseases / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D061987:Sf9 Cells; D018411:Spodoptera
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D015707:HN Protein; D011994:Recombinant Proteins
keywords: Newcastle disease virus; antibody detection; hemagglutination inhibition; hemagglutinin-neuraminidase protein